Apoptosis of bladder transitional cell carcinoma T24 cells induced by adenovirus-mediated inducible nitric oxide synthase gene transfection

作者：Tan, Jing; Zeng, Qing^*; Jiang, Xian-Zheng; He, Le-Ye; Wang, Jin-Rong; Yao, Kun; Wang, Chang-Hui

来源：Chinese Journal of Cancer Research, 2013, 25(5): 593-599.

DOI：10.3978/j.issn.1000-9604.2013.10.11

• Bladder cancer
• T24 cell line
• iNOS gene
• gene therapy

Objectives: To investigate the effects of adenovirus-mediated inducible nitric oxide synthase gene transfection on bladder transitional cell carcinoma T24 cells, and to provide novel insights and approaches to clinical therapies against bladder transitional cell carcinoma.
Methods: Firstly, construct recombinant adenovirus vector pAd-iNOS of iNOS, followed by transfection of pAd-iNOS into HECK293 packaging cells. Thirdly, harvest recombinant adenovirus rAd-iNOS after amplification and purification procedures Finally, transfect the recombinant adenovirus rAd-iNOS into human bladder carcinoma T24 cells and examine the effect of rAd-iNOS transfection on apoptosis of T24 and possible mechanism.
Results: As shown by this study, the recombinant adenovirus rAd-iNOS was constructed successfully. The virus titer was 5.8x10(8) PFU/mL and recombinant was verified by PCR analysis. Transfection of adenovirus rAd-iNOS into T24 cells could induce secretion of high NO concentration, P53 protein expression up-regulation, as well as promotion of T24 cell apoptosis.
Conclusions: The transfection of human bladder carcinoma T24 cells from recombinant adenovirus rAd-iNOS was confirmed to induce intracellular iNOS over-expression, high production of NO, up-regulation of intracellular P53 expression and promotion of cell apoptosis.

• 出版日期2013-10
• 单位中南大学